The current available molecular method to detect infectious bursal disease virus (IBDV) is by reverse transcriptase-polymerase chain reaction (RT-PCR). However, the conventional PCR is time consuming, prone to error and less sensitive. In this study, the performances of Sybr Green I real-time PCR, enzyme-linked immunosorbent assay (ELISA) and conventional agarose detection methods in detecting specific IBDV PCR products were compared. We found the real-time PCR was at least 10 times more sensitive than ELISA detection method with a detection limit of 0.25pg. The latter was also at least 10 times more sensitive than agarose gel electrophoresis detection method. The developed assay detects both very virulent and vaccine strains of IBDV but not other RNA viruses such as Newcastle disease virus and infectious bronchitis virus. Hence, Sybr Green I-based real-time PCR is a highly sensitive assay for the detection of IBDV.

Introduction: Hepatitis C virus (HCV) genotyping is very important for the clinical management of HCVinfected patients. The aim of this study was to determine the genotypes of HCV-infected patients and to identify their risk factors and comorbidities. Materials and Methods: This was an observational, cross sectional study in which forty (40) HCV-infected patients attending Gastroenterology Clinic, Hospital Tengku Ampuan Afzan (HTAA) Kuantan Pahang were recruited for the study, from January to July 2014. Nucleotide sequence analysis of the 5’UTR and NS5B region were performed to identify the viral genotypes. Results: Of the 40 samples, 31 (77.5%) isolates were successfully classified into their genotypes and subtypes; 3a (57.5%), 1a (12.5%), 3b (2.5%) and 1b (2.5%). No genotype 2, 4, 5 and 6 were found in this study. However, there was one mixed-genotype (3a/1a) HCV infection. Risk factors and co-morbidities found in this study include IVDUs, haemodialysis, blood transfusion, surgery and co-infection with HIV. Conclusion: Genotype 3 followed by genotype 1 were the common HCV genotypes found in this study population. Furthermore, the highest risk factors and co-morbidities were IVDUs and co-infection with HIV.

Background: Human enterovirus 71 (HEV71) is a common aetiology of hand-foot-and-mouth-disease (HFMD) where it occasionally causes CNS illnesses. In 1997, Sarawak experienced a HEV71-associated HFMD outbreak with 34 deaths reported. Following the outbreak, a prospective HFMD surveillance study showed that HE71-associated HFMD outbreaks occurred in Sarawak every three years, each caused predominantly by different genogroups of HEV71; genogroup-B3 in 1997, genogroup-B4 in 2000, and genogroup-B5 in 2003 This study attempts to investigate the circulation of HEV71 in between the three-year cycle outbreaks before an expected outbreak in 2006 in Sarawak.Methods: Three communities, Villages-A, B and C, were selected based on their locations with reference to the Rajang river, the epicenter of the 1997 outbreak. Stool samples were collected through 2 visits; the first collection in mid-2005 and the second in February 2006. Filter-sterilized samples were inoculated into rhabdomyosarcoma and QB1-293A cells. Virus cultures were observed for cytopathic effect (CPE) daily for 14-days. Upon blind-passage, cultures without CPE were considered negative. Reverse-transcription-polymerase-chain-reaction (RT-PCR) on positive cultures was done using primers for partial VP1 previously described by Oberste et al. RT-PCR positive products were subjected to DNA sequencing and molecular analysis.Results: During the first collection, 106 samples were collected from Village-A, 428 from Village-B and 183 from Village-C. The second collection yielded 88 samples from Village-A, 410 from Village-B and 177 from Village-C. While, no virus was isolated from the first collection of Village-A, 4% of samples from Village-B and 6% of samples from Village-C yielded enteroviruses respectively. From the second collection, 8% of samples from Village-A, 6.8% from Village-B and 4.5% from Village-C, yielded enteroviruses. The viruses isolated were identified mostly as echovirus-6 (E6), E7, E19, coxsackievirus-A (CA) 20, CA21, CA24 and coxsackievirus-B (CB) 4.Conclusion: HEV71 was not circulating in sampled villages but enteroviruses species B and C were.